• The Journal of Dong Guk Oriental Medicine
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• v.9
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• pp.179-191
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• 2000

Anti-bone resorption properties of the Korean herbal medicine, CEDR, which is comprised 5 herbs of [Drynariae Rhizoma, Loranthi Ramus, Cibotii Rhizoma, Amydae carapax, Psoraleae semen], were investigated. Mouse calvarial osteoblast cells were isolated and cultured. Mouse osteoblasts, which were stimulated by PTH, $1,25(OH)_2D_3$, $TNF-\alpha$ and IL-1 as bone resorption agents, showed increased collagenolysis by producing the active gelatinase. IL-1 in stimulating bone resorption was examined using fetal mouse long bone organ culture. IL-1 stimulated bone resorption and produced marked resorption when present simultaneously. The results of in vitro cytotoxicities showed that CEDR extracts have no any cytotoxicities in concentrations of $1-60{\mu}g/ml$ and furthermore there is no any cytotoxicity even in concentration of $120{\mu}g/ml$ on mouse calvarial bone cells. CEDR extracts had protective activity against PTH (5 units/ml, or $IL-1{\alpha}$ (1 ng/ml) or $TNF-\alpha$ or $1,25(OH)_2D_3$ (20 ng/ml), $IL-1{\alpha}$ and $IL-1{\beta}-induced$ collagenolysis in the mouse calvarial cells. Pretreatment of the CEDR extracts for 1 h, which by itself had little effect on cell survival, did not enhance the collagenolysis, nor significantly reduced the collagenolysis by pretreatment. Furthermore, the medicinal extracts were shown to have the protective effects against collagenolysis induced by $IL-1{\alpha}$ and $IL-1{\beta}$. Pretreatment of the extracts for 1 h significantly reduced the collagenolysis. Interestingly, the CEDR extracts were shown to have the inhibiting effects against gelatinase enzyme and processing activity induced by the bone resorption agents of PTH, $1,25(OH)_2D_3$, $TNF-\alpha$, $IL-1{\beta}$ and $IL-1{\alpha}$ with strong protective effect in pretreatment with the extracts. CEDR extracts were shown to have the inhibiting effects against $IL-1{\alpha}-$ and $IL-1{\beta}-stimulated$ bone resorption and the effect of the pretreatment with a various concentrations of the medicinal extracts were significant. These results indicated that the CEDR extracts are highly stable and applicable to clinical uses in osteoporosis.

• Journal of Life Science
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• v.16 no.2 s.75
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• pp.315-322
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• 2006

FS11052, a novel microbial metabolite from Streptomyces spp. was identified as a small molecular substance and shown inhibition activities for the release of neurotransmitter from rat hippocampal neuron and PC12 cells. FS11052 is an inhibitor of tritiated norepinephrine ($[^3H]-NE$) release in high $K^+$ buffer solution containing ionomycin, indicating that FS11052 inhibits neurotransmitter release after the influx of $Ca^{2+}$ ions. When examined the effect of FS11052 on glucuronidase release from guinea pig neutrophils, FS11052 inhibited glucuronidase release: when treated with $5{\mu}g/ml$ of FS11052, which was not induced cellular cytotoxicity. The fact that the glucuronidase release in neutrophil and norepinephrine release in neuron was inhibited suggests the similarity in the locations and the mechanisms of FS11052 action targets. When treated with $5{\mu}g/ml$ of FS11052, $[^3H]-NE$ release and neurite extension for both rat hippocampal neurons and PC12 cells were prevented. These observations of FS11052 functioning as an inhibitor of neurotransmitter release suggest that FS11052 has an important role in synaptic transmission in neuron.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.11
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• pp.1552-1563
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• 2016

To examine the cognitive function of onion (Allium cepa L.) beverages (odourless and fortified), we analyzed in vitro neuronal cell protection against $H_2O_2$-induced cytotoxicity and performed in vivo tests on amyloid beta ($A{\beta}$)-induced cognitive dysfunction. Cellular oxidative stress and cell viability were evaluated by DCF-DA assay and MTT assay. These results show that fortified beverage resulted in better neuronal cell protection than odourless beverage at lower concentration ($0{\sim}100{\mu}g/mL$). Fortified beverage also showed more excellent acetylcholinesterase (AChE) inhibitory activity ($IC_{50}$: 4.20 mg/mL) than odourless beverage. The cognitive functions of odourless beverage and fortified beverage in $A{\beta}$-induced neurotoxicity were assessed by Y-maze, passive avoidance, and Morris water maze tests. The results show improved cognitive function in both groups treated with beverages. After in vivo tests, cholinergic activities were determined based on AChE inhibition and acetylcholine levels, and antioxidant activities were measured as SOD, oxidized glutathione (GSH)/total GSH ratio, and MDA levels in mouse brain tissue. In a Q-TOF UPLC/MS system, main compounds were analyzed as follows: odourless beverage (five types of sugars and three types of phenolics) and fortified beverages (six types of phenolics and two types of steroidal saponins).

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.6
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• pp.809-815
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• 2015

In this study, we confirmed biological compounds from methanol (MeOH) extract of processed Polygoni Multiflori Radix (PPMR), and the radical scavenging effect and oxidative stress protective activity of MeOH extract of PPMR were investigated under in vitro conditions using LLC-$PK_1$ renal epithelial cells. In HPLC analysis, MeOH extract of PPMR contained four species of biological compounds named 2,3,5,4'-tetrahydroxystilbene 2-O-${\beta}$-D-glucoside, emodin, chrysophanol, and rhein. 2,3,5,4'-Tetrahydroxystilbene 2-O-${\beta}$-D-glucoside was detected as the main compound in PPMR as 115.02 mg/kg. MeOH extract of PPMR showed 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) diammonium salt (ABTS), and hydroxyl radical scavenging activities in a concentration- dependent manner. In particular, upon $50{\mu}g/mL$ of PPMR extract treatment, DPPH, ABTS, and hydroxyl radical scavenging activities were approximately 48.4%, 57.9%, and 81.2%, respectively. LLC-$PK_1$ cell viability declined in response to oxidative stress induced by pyrogallol, sodium nitroprusside (SNP), and morpholinosydnonimine (SIN-1) generators of NO, $O_2{^-}$, and $ONOO^-$, respectively. However, MeOH extract of PPMR significantly and dose-dependently inhibited oxidative-stressed LLC-$PK_1$ cell cytotoxicity. In fact, upon $50{\mu}g/mL$ of PPMR extract treatment, LLC-$PK_1$ cell viabilities were approximately 82.1%, 89.1%, and 77.6% compared to stress levels induced by pyrogallol, SNP, and SIN-1, respectively.

• Journal of Radiopharmaceuticals and Molecular Probes
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• v.9 no.1
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• pp.9-16
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• 2023

Liposomes as drug delivery system have proved useful carrier for various disease, including cancer. In addition, perfluorocarbon cored microbubbles are utilized in conjunction with high-intensity focused-ultrasound (HIFU) to enable simultaneous diagnosis and treatment. However, microbubbles generally exhibit lower drug loading efficiency, so the need for the development of a novel liposome-based drug delivery material that can efficiently load and deliver drugs to targeted areas via HIFU. This study aims to develop a liposome-based drug delivery material by introducing a substance that can burst liposomes using ultrasound energy and confirm the ability to target tumors using PET imaging. Liposomes (Lipo-DOX, Lipo-DOX-Au, Lipo-DOX-Au-RGD) were synthesized with gold nanoparticles using an avidin-biotin bond, and doxorubicin was mounted inside by pH gradient method. The size distribution was measured by DLS, and encapsulation efficiency of doxorubicin was analyzed by UV-vis spectrometer. The target specificity and cytotoxicity of liposomes were assessed in vitro by glioblastoma U87mg cells to HIFU treatment and analyzed using CCK-8 assay, and fluorescence microscopy at 6-hour intervals for up to 24 hours. For the in vivo study, U87mg model mouse were injected intravenously with 1.48 MBq of ⁶⁴Cu-labeled Lipo-DOX-Au and Lipo-DOX-Au-RGD, and PET images were taken at 0, 2, 4, 8, and 24 hours. As a result, the size of liposomes was 108.3 ± 5.0 nm at Lipo-DOX-Au and 94.1 ± 12.2 nm at Lipo-DOX-Au-RGD, and it was observed that doxorubicin was mounted inside the liposome up to 52%. After 6 hours of HIFU treatment, the viability of U87mg cells treated with Lipo-DOX-Au decreased by around 20% compared to Lipo-DOX, and Lipo-DOX-Au-RGD had a higher uptake rate than Lipo-DOX. In vivo study using PET images, it was confirmed that ⁶⁴Cu-Lipo-DOX-Au-RGD was taken up into the tumor immediately after injection and maintained for up to 4 hours. In this study, drugs released from liposomes-gold nanoparticles via ultrasound and RGD targeting were confirmed by non-invasive imaging. In cell-level experiments, HIFU treatment of gold nanoparticle-coupled liposomes significantly decreased tumor survival, while RGD-liposomes exhibited high tumor targeting and rapid release in vivo imaging. It is expected that the combination of these models with ultrasound is served as an effective drug delivery material with therapeutic outcomes.

• Food Science and Preservation
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• v.30 no.6
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• pp.1095-1106
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• 2023

This study investigated the anti-oxidative and anti-inflammatory effects of Aster glehni (AG) extract in RAW 264.7 cells and Caenorhabditis elegans. The total polyphenol and flavonoid contents were higher in the ethanol extracts than in the hot water extracts. As a result of measuring the moisture contents (%) and extraction yields (%) of AG and drying A. glehni for processing (DAG), 70% ethanol, which has the highest percentage of extraction yield, was selected as the final solvent. DPPH radical scavenging activity showed higher antioxidant activity of ethanol extracts of DAG than AG. The cytotoxicity assay of the AG or DAG ethanol extracts was treated at different concentrations (25, 50, and 100 ㎍/mL), and cell viability rates were higher than 80% at all concentrations. The LPS-stimulated nitric oxide (NO) production in RAW 264.7 was significantly reduced at all concentrations of AG and DAG groups. As a result of measuring the gene expression of iNOS, which induces NO production, the AG or DAG group decreased by 33% and 32%, compared with the phosphate buffer saline (PBS) group. Under inflammatory stress conditions, the survival rate of C. elegans treated with AG or DAG ethanol extract with LPS showed concentration-dependent improvement in survival rate compared with the PBS group. Considering these results, AG could potentially be developed as an antioxidant and anti-inflammatory functional food material.

• The Korean Journal of Mycology
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• v.26 no.2 s.85
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• pp.246-255
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• 1998

Ganoderan (GAN), an immunomodulating ${\beta}-glucan$ of G. lucidum, induces potent antitumor immunity in tumor-bearing mice. This study was set up to elucidate the ability of macrophage activation of GANs. GAN-treated Raw 264.7 macrophages showed enhanced production of nitric oxide (NO). The ability of GANs to produce NO was based on differences in chemical composition of GANs obtained from the mycelium on various carbon sources and mycelial fractionation. The highest NO production was observed in CW-AS-WS polysaccharide which was extracted from the mycelial wall. GAN-treated Raw 264.7 cells gave a 2-to 5-fold (24 hr) formation of NO levels compared with those treated with medium only. Partial removal of the protein in the extracellular GAN by TCA treatment did appreciably reduce its capacity to secrete NO. The mixture effect of GAN and LPS increased the nitric oxide secretion from RAW 264.7. The cell proliferation of GAN-treated Raw 264.7 cell tines inhibited as compared with its control. Of the culture supernatant of macrophage activated by GAN, the percentage of cytotoxicity against mouse leukemia L1210 cells was slightly dependent on the amount of NO in the culture supernatants of the activated-macrophages. These results indicate that the ${\beta}-glucan-related$ polysaccharides of the higher fungus activate macrophage and release nitric oxide. It also suggests that murine macrophages possess certain receptors for ${\beta}-anomeric$ glucans and play a critical role of ${\beta}-glucan-related$ tumor killing mechanism.

• Food Science and Preservation
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• v.24 no.8
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• pp.1149-1157
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• 2017

Inflammation is the first response of the immune system to infection or irritation in our body. The use of medicinal plants has been widely applied as an alternative source for drug development. One of marine natural resources, the anti-inflammatory effect of Ishige sinicola ethanol extract (ISEE), was evaluated by using LPS-induced RAW 264.7 cell and mice model. As a result, the production of nitric oxide (NO) and pro-inflammatory cytokines (IL-6, IL-$1{\beta}$, TNF-${\alpha}$) were inhibited with increasing concentration of ISEE without any cytotoxicity. Furthermore, ISEE suppressed the expression of not only inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-${\kappa}B$) p65, and mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK) 1/2, p38, and c-Jun N-terminal kinase (JNK) in a dose-dependent manner. In mice ear edema test, the formation of edema was reduced at the highest dosage of ISEE and the reduction of the number of infiltrated mast cells was observed in histological analysis. These results indicate that ISEE has a potent anti-inflammatory activity and can be used as a pharmaceutical material for many kinds of inflammatory disease.

• Journal of Food Hygiene and Safety
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• v.38 no.6
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• pp.430-441
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• 2023

Velvet antler is widely used as a traditional medicine, and numerous studies have demonstrated its tremendous nutritional and medicinal values including immunity-enhancing effects. This study aimed to investigate different deer velvet extracts (Sample 1: raw extract, Sample 2: dried extract, and Sample 3: freeze-dried extract) for proximate composition, uronic acid, sulfated glycosaminoglycan, sialic acid, collagen levels, and chemical components using ultra-performance liquid chromatography-quadrupole-time-of-light mass spectrometry. In addition, we evaluated the cytotoxic effect of the deer velvet extracts on BV2 microglia, HT22 hippocampal cells, HaCaT keratinocytes, and RAW264.7 macrophages using the cell viability MTT assay. Furthermore, we evaluated acute toxicity of the deer velvet extracts at different doses (0, 500, 1000, and 2000 mg/kg) administered orally to both male and female ICR mice for 14 d (five mice per group). After treatment, we evaluated general toxicity, survival rate, body weight changes, mortality, clinical signs, and necropsy findings in the experimental mice based on OECD guidelines. The results suggested that in vitro treatment with the evaluated extracts had no cytotoxic effect in HaCaT keratinocytes cells, whereas Sample-2 had a cytotoxic effect at 500 and 1000 ㎍/mL on HT22 hippocampal cells and RAW264.7 macrophages. Sample 3 was also cytotoxic at concentrations of 500 and 1000 ㎍/mL to RAW264.7 and BV2 microglial cells. However, the mice treated in vivo with the velvet extracts at doses of 500-2000 mg/kg BW showed no clinical signs, mortality, or necropsy findings, indicating that the LD50 is higher than this dosage. These findings indicate that there were no toxicological abnormalities connected with the deer velvet extract treatment in mice. However, further human and animal studies are needed before sufficient safety information is available to justify its use in humans.

• KSBB Journal
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• v.24 no.4
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• pp.327-333
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• 2009

The antioxidant and anti-inflammatory activities of ethanol extract of Malus micromalus were studied in vitro. Ethanol extract of M. micromalus showed scavenging effects on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and nitric oxide (NO) radicals. In addition, ethanol extract of M. micromalus inhibited the generation of superoxide anion ($O_2^-$) radical and uric acid by xanthine oxidase. We also investigated the effect of ethanol extract of M. micromalus on NO production in a lipopolysaccharide (LPS)-stimulated murine macrophage RAW 264.7 cells. Ethanol extract of M. micromalus significantly inhibited NO production and this inhibitory action was not due to the cytotoxicity. The expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) was markedly down-regulated by ethanol extract of M. micromalus. These results indicate that the inhibitory action of ethanol extract of M. micromalus on NO production in LPS-stimulated macropages might be due in part to abrogation of iNOS and COX-2 protein induction. Taken together, this study suggests that ethanol extract of M. micromalus could contribute to the chemoprevention and therapy of oxidative stress and inflammation.